Difference gel electrophoresis using matched multiple dyes

Electrophorese differentielle sur gel faisant appel a des colorants multiples apparies

Abstract

A process and a kit are provided for detecting differences in two or more samples of protein. Protein extracts are prepared, for example, from each of a different group of cell samples (1, 2) to be compared. Each protein extract is labeled with a different one of a luminescent dye from a matched set of dyes. The matched dyes have generally the same ionic and pH characteristics but emit light at different wavelengths to exhibit a different color uponluminescence detection. The labeled protein extracts are mixed together and electrophoresed together. The gel (4) is observed to detect proteins unique to one sample or present in a greater ratio in one sample than in the other. Those unique or excess proteins will fluoresce the color of one of the dyes used. Proteins common to each sample migrate together and fluoresce the same.
Cette invention concerne un procédé ainsi qu'un nécessaire permettant de détecter des différences entre deux échantillons de protéines ou plus. Des extraits de protéines sont préparés, par exemple, à partir de chaque groupe différent d'échantillons (1, 2) de cellules à comparer. Chaque extrait de protéines est marqué avec un colorant luminescent différent provenant d'un ensemble de colorants appariés. Ces colorants appariés possèdent, de manière générale, les mêmes caractéristiques ioniques et de pH, mais émettent de la lumière à des longueurs d'ondes différentes de manière à produire différentes couleurs lors de la détection de luminescence. Les extraits de protéines marqués sont mélangés entre eux et soumis ensemble à l'électrophorèse. Le gel (4) est ensuite observé afin de détecter des protéines propres à un seul échantillon, ou des protéines présentes dans un échantillon dans une plus grande proportion que dans l'autre. Ces protéines propres ou en quantité excessive émettront une fluorescence de la couleur de l'un des colorants employés. Les protéines communes à chaque échantillon migrent ensemble et émettent la même fluorescence.

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